Detection and occurrence of waterborne bacterial and viral pathogens

Black, E. Kathleen; Finch, Gordon R.

doi:10.1002/j.1554-7531.1993.tb00050.x

Cited by 9 publications

(3 citation statements)

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“…1999). The possible contamination of reconditioned wastewater by E. coli O157:H7 and its use for agricultural purposes pose a potential for disease outbreaks (Black and Finch 1993). The aim of our work was to verify the suitability and utility of a rapid semi‐automated real‐time PCR assay for detection of E. coli O157:H7 in dairy wastewater.…”

Section: Discussionmentioning

confidence: 99%

“…Reclamation and reuse of wastewater in agricultural processes are often used as means for extending water supplies and decreasing water fees (Rajkowski and Rice 1999). Although methods for the effective treatment of wastewater exist and are essential to ensure the safety of drinking water and crops, the wastewater originated from dairy operation may harbour different bacterial species including human pathogens such as enterohaemorrhagic Escherichia coli O157:H7 (Black and Finch 1993; Ibekwe et al. 2002).…”

Section: Introductionmentioning

confidence: 99%

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Real-time PCR for the detection of Escherichia coli O157:H7 in dairy and cattle wastewater

Spano

Beneduce

Terzi

et al. 2005

Lett Appl Microbiol

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Aims: Developing and evaluating a rapid real-time polymerase chain reaction (PCR) method for the identification of Escherichia coli O157:H7 in cattle and dairy wastewater samples produced from mozzarella cheese factories, without pre-enrichment step before DNA extraction. Methods and Results: Wastewater samples were collected from a dairy farm producing mozzarella cheese and located in Puglia (south of Italy). Plate count and other microbial assays were performed 1 h after sampling. Wastewater samples were artificially inoculated with 10 4 , 10 7 and 10 8 cells ml )1 of E. coli O157:H7, strain EDL933.PCR protocols for stx1, stx2 and eae genes were first tested on pure DNA extracted from type strains, in order to optimize the amplification conditions and reagent concentration before real-time PCR experiments. Three specific fragments of ca 106, 150 and 200 bp corresponding to genes eae, stx1 and stx2, respectively, were obtained. Real-time PCR experiments were performed with DNA extracted from dairy and manure wastewater samples inoculated with 10 4 , 10 7 and 10 8 colony-forming units (CFU) ml )1 of E. coli O157:H7 strain EDL 933. The sensitivity limit of the assay was 10 )1 pg ll )1 for eae, stx2 and 16SrRNA, and 1 pg ll )1 for stx1 gene respectively. Conclusions:A real-time PCR protocol has been developed and used in order to identify potential pathogens in dairy wastewater, in which previous methods (including standard PCR) failed to work. Significance and Impact of the Study: Cattle and dairy wastewater samples produced from mozzarella cheese factories may harbour verocytotoxin-producing E. coli. The availability of rapid and sensitive molecular methods may be useful to monitor the persistence of verocytotoxin-producing E. coli in general and to assess the effectiveness of wastewater treatments.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Real-time PCR for the detection of Escherichia coli O157:H7 in dairy and cattle wastewater

Spano

Beneduce

Terzi

et al. 2005

Lett Appl Microbiol

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“…Although most strains of E. coli are not pathogenic, they can act as opportunistic pathogens causing infections in immune‐compromised hosts. In the E. coli family are also present pathogenic strains that when ingested can cause gastrointestinal illness in healthy humans 3, 4…”

Section: Introductionmentioning

confidence: 99%

SYBR green real time‐polymerase chain reaction as a rapid and alternative assay for the efficient identification of all existing Escherichia coli biotypes approved directly in wastewater samples

Chetta

Bafunno

Grillo³

et al. 2012

Biotechnology Progress

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Escherichia coli has been recognized as the principal indicator of fecal contamination of water. Indeed, E. coli is the only species in the coliform group found in relationship with gastrointestinal tract of human and warm‐blooded animals and subsequently excreted in large numbers in the human feces. To obtain a complete picture of water quality and therefore, a better protection of public health, different techniques for water analysis have been proposed. In this article, we describe an alternative method that uses SYBR green real time‐polymerase chain reaction (RT‐PCR) technology to identify and quantify all E. coli biotypes in a group of wastewater samples collected from a wastewater depurator located in South of Italy. This new RT‐PCR protocol is accurate in measuring the concentration of chromosomal E. coli DNA using the amplification of three new specific fragments of the following bacteria genes: CadC, HNS, and Allan whose sequence is specific for E. coli family and conserved in all E. coli subtypes. This method allowed us to detect the presence of all E. coli biotypes directly in wastewater samples and estimated the correspondence between colony forming units and bacterial DNA concentrations. The availability of a rapid and sensitive method may be useful to monitor the persistence of E. coli in water, to evaluate the efficiency of wastewater purification treatments and the possible recycle for agricultural use. Furthermore, the development of a simple and routine method to monitor water quality with RT‐PCR analysis can encourage the testing of a higher number of samples. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 28: 1106–1113, 2012

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